Direct IgG epitope mapping on bacterial AB toxins by cryo-EM

被引:0
|
作者
Nguyen, Tri [1 ]
Song, Jeongmin [1 ]
机构
[1] Cornell Univ, Dept Microbiol & Immunol, Ithaca, NY 14853 USA
来源
STAR PROTOCOLS | 2021年 / 2卷 / 04期
关键词
Antibody; Cryo-EM; Immunology; Microbiology; Microscopy; Protein Biochemistry; Structural Biology;
D O I
10.1016/j.xpro.2021.100852
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
This cryo-EM protocol was used to determine the B cell epitope map on the CdtB subunit of typhoid toxin, an A2B5 toxin secreted by Salmonella Typhi during infection. Immunoglobulin G (IgG) was directly mixed with typhoid toxin in this protocol, different from our previous cryo-EM protocol that uses the Fab fragments in place of IgG. This simple approach requires smaller amounts of materials, supporting the broader use of this protocol for determining antibody recognition sites on various antigens. For complete details on the use and execution of this protocol, please refer to Ahn et al. (2021) and Nguyen et al. (2021).
引用
收藏
页数:15
相关论文
共 50 条
  • [31] Noise models and cryo-EM drift correction with a direct-electron camera
    Shigematsu, H.
    Sigworth, F. J.
    ULTRAMICROSCOPY, 2013, 131 : 61 - 69
  • [32] Cryo-EM structure of the bacterial intramembrane metalloprotease RseP in the substrate-bound state
    Asahi, Kikuko
    Hirose, Mika
    Aruga, Rie
    Shimizu, Yosuke
    Tajiri, Michiko
    Tanaka, Tsubasa
    Adachi, Yuriko
    Tanaka, Yukari
    Kaneko, Mika K.
    Kato, Yukinari
    Akashi, Satoko
    Akiyama, Yoshinori
    Hizukuri, Yohei
    Kato, Takayuki
    Nogi, Terukazu
    Science Advances, 2025, 11 (09)
  • [33] New Opportunities Created by Single-Particle Cryo-EM: The Mapping of Conformational Space
    Frank, Joachim
    BIOCHEMISTRY, 2018, 57 (06) : 888 - 888
  • [34] Cryo-EM structures reveal multiple stages of bacterial outer membrane protein folding
    Doyle, Matthew Thomas
    Jimah, John R.
    Dowdy, Tyrone
    Ohlemacher, Shannon, I
    Larion, Mioara
    Hinshaw, Jenny E.
    Bernstein, Harris D.
    CELL, 2022, 185 (07) : 1143 - +
  • [35] THE SUPRAMOLECULAR STRUCTURE OF THE BACTERIAL STRESSOSOME REVEALED BY CRYO-EM UNVEILS ITS MECHANISM OF ACTIVATION
    Williams, Allison
    PROTEIN SCIENCE, 2019, 28 : 154 - 154
  • [36] Cryo-EM investigation of the subconductance state of ryanodine receptor calcium release channels induced by scorpion toxins
    Van Petegem, Filip
    Haji-Ghassemi, Omid
    Chen, Yu
    Woll, Kellie
    Valdivia, Carmen R.
    Gurrola, Georgina B.
    Cai, Wenxuan
    Li, Songhua
    Valdivia, Hector H.
    BIOPHYSICAL JOURNAL, 2023, 122 (03) : 164A - 164A
  • [37] Cryo-EM structure of bacterial nitrilase reveals insight into oligomerization, substrate recognition, and catalysis
    Aguirre-Sampieri, Sergio
    Casanal, Ana
    Emsley, Paul
    Garza-Ramos, Georgina
    JOURNAL OF STRUCTURAL BIOLOGY, 2024, 216 (02)
  • [38] Validation of cryo-EM structures of SARS-CoV-2 and mapping genomic mutations
    Malhotra, Sony
    Winn, Martyn
    Joseph, Agnel Praveen
    ACTA CRYSTALLOGRAPHICA A-FOUNDATION AND ADVANCES, 2021, 77 : C615 - C615
  • [39] Fabrication of carbon films with ∼500 nm holes for cryo-EM with a direct detector device
    Marr, Chelsea R.
    Benlekbir, Samir
    Rubinstein, John L.
    JOURNAL OF STRUCTURAL BIOLOGY, 2014, 185 (01) : 42 - 47
  • [40] Direct calculation of cryo-EM and crystallographic model maps for real-space refinement
    Urzhumtsev, Alexandre G.
    Urzhumtseva, Ludmila M.
    Lunin, Vladimir Y.
    ACTA CRYSTALLOGRAPHICA SECTION D-STRUCTURAL BIOLOGY, 2022, 78 : 1451 - 1468